title: Expression, purification, crystallization and preliminary X-ray analysis of an archaeal protein homologous to plant nicotianamine synthase.
abstract: In plants, nicotianamine synthase (NAS) plays a key role in metal homeostasis as it catalyzes the formation of nicotianamine, an important iron and nickel chelator and a precursor of plant phytosiderophores. Here, the crystallization of a protein from Methanothermobacter thermoautotrophicus (MTH675; referred to here as MtNAS) that appears to be homologous to plant NAS is reported. Purification of this protein showed a monomer-dimer equilibrium that could be displaced by using a reducing agent such as DTT. Crystals belonging to space group P2(1)2(1)2(1) and containing dimers of MtNAS were grown by the vapour-diffusion method using polyethylene glycol 3350 as precipitant. A complete native X-ray data set was collected to 1.7 A resolution at a synchrotron source.
authors.affiliation: CEA, DSV, IBEB, Laboratoire de Bioénergétique Cellulaire, 13108 Saint Paul Lez Durance, France.
authors.forename: Cyril
authors.initials: C
authors.lastname: Dreyfus
authors.forename: David
authors.initials: D
authors.lastname: Pignol
authors.forename: Pascal
authors.initials: P
authors.lastname: Arnoux
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D019843:Archaeal Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D002850:Chromatography, Gel; D003460:Crystallization; D018360:Crystallography, X-Ray; D017931:DNA Primers; D017013:Methanobacteriaceae / Q000737:chemistry* / Q000235:genetics; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics; D016133:Polymerase Chain Reaction
chemical_list: D019843:Archaeal Proteins; D017931:DNA Primers; D010940:Plant Proteins; D019883:Alkyl and Aryl Transferases; C118230:nicotianamine synthase